[Summary] Thoracic paravertebral block ( TPVB) can offer intra-postoperative analgesia for thoracic , cardiac, and breast operations .In recent years , the development of ultrasonic technology provides a platform for real-time and visual never block , which can further improve the success rate and reduce the incidence of complications .In this article, we reviewed the various approaches of ultrasound guidance for thoracic paravertebral blockade , and explored the latest progress of different technologies .

OBJECTIVE:To determine camphor in compound menthol nose drop by RP-HPLC.METHODS:Waters No-va-pak C 18 was taken as the chromatographic column,the mobile phase was composed of chloroform-methanol-water(14∶52∶30)with a flow speed at1.0ml/min and detection wavelength at289nm,the column temperature was35℃.RESULTS:The linear range for camphor was0.8～5.6?g/ml(r=0.9998)and the average recovery rate was99.4%(RSD=0.7%).CONCLUSION:The method is specific,convenient and accurate,which can be used for the quality control of the preparation.

Objective To study some sensitive electrophysiological parameters in surveillance of allograft rejection.Methods Forty rats underwent heterotopic heart transplantations.IMEG was re- corded by an epicardiac unipolar pacing lead fixed at the right ventricular outflow tract.QRS amplitude and heart rate were determed daily in 10 syngeneic and 30 allogeneic transplants.Syngeneic transplants were killed at 7 th postoperative day,and allogeneic transplants killed at 3 rd,5 th and 7 th postopera- tive day.Histopathologie studies were performed at every transplanted heart.Results In syngeneic group,QRS amplitude kept constant after the transplantation while no significant differences were ob- served at the 3 rd,5 th and 7 th postoperative day.QRS amplitude was dropped obviously in allogeneic group after the first two postoperative days whereas significant differences were observed at the rejec- ting and non-rejecting hearts.Conclusions IMEG is a valid method to monitor acute allograft rejec- tion.QRS amplitude is a more sensitive electrophysiological parameter to diagnose severe rejections than heart rate,while mild rejections were not detected by this method.

Six patients with ST segment elevated acute myocardial infarction (AMI), who were 52.5 years old in average, were enrolled and performed the treatment at Tongren Hospital from November 2003 to June 2004. Following percutanecus transluminal coronary angioplasty and stent revascularization, autologous bone marrow stem cell (BMSC) transplantation was performed after informed consent was obtained. Patients were subcutaneously injected with granulocyte colony-stimulating factor (G-CSF) at 1 week before transplantation. When CD34+ cells going up to 1%-3% in peripheral blood, mononuclear cells in peripheral blood were harvested,purified, and further infused into the infarcted related coronary artery with an over-the-wire balloon catheter. Following up was performed every half a year. Four years later, the infarcted area of these patients was further decreased by 8.03%, in the basic descent of 42.7% at 3 months averagely; total infracted area descent was 50.73%, but ejection fraction increased by 4.6% from 50.8%. There was no serious coronary artery restenosis and/or stenosis formation which need revascularization upon angiography.

To study the cartilage differentiation of mouse mesenchymal stem cells (MSCs) induced by cartilage-derived morphogenetic proteins-2 in vitro, the MSCs were isolated from mouse bone marrow and cultured in vitro. The cells in passage 3 were induced into chondrogenic differentiation with different concentrations of recombinant human cartilage-derived morphogenetic proteins-2 (0, 10, 20, 50 and 100 ng/mL). After 14 days of induction, morphology of cells was observed under phase-contrast microscope. Collagen II mRNA and protein were examined with RT-PCR, Western blotting and immunocytochemistry respectively and the sulfate glycosaminoglycan was measured by Alcian blue staining. RT-PCR showed that CDMP-2 could promote expression of collagen II mRNA in an dose-dependant manner, especially at the concentration of 50 ng/mL and 100 ng/mL. Immunocytochemistry and Western blotting revealed a similar change. Alcian blue staining exhibited deposition of typical cartilage extracellular matrix. Our results suggest that mouse bone marrow mesenchymal stem cells can differentiate into chondrogenic phonotype with the induction of CDMP-2 in vitro, which provides a basis for further research on the role of CDMP-2 in chondrogenesis.
Bone Marrow Cells/*cytology ; Bone Morphogenetic Proteins/*pharmacology ; Cell Differentiation/*drug effects ; Cells, Cultured ; Chondrocytes/*cytology ; Chondrogenesis/drug effects ; Chondrogenesis/physiology ; Mesenchymal Stem Cells/*cytology ; Recombinant Proteins/pharmacology

Objective To establish RT-PCR-RFLP method for studying the genotype of wild mea-sles virus strains isolated from Tianjin area from 2002 to 2008. Methods Isolations of measles virus were carried out by tissue culture method from urine and throat swab specimens collected from suspected cases. RNA were extracted from the virus specimens. The 594 bp fragment of C terminal of the N (nucleoprotein) gene was amplified by one-step RT-PCR, then the PCR products were digested with Bcn I , separated on agarose gel electrophoresis and then analyzed by the method of RFLP (restriction fragment length polymor-phism). In addition, above results were compared with DNA sequencing. Phylogenetic tree was plotted based on the results for the genetic relationship and distance analysis. Results Sixty-nine measles virus strains were isolated from 189 specimens from 2002 to 2008, of which the C terminals of N gene were all de-tected positive. Among the 69 strains of measles virus isolates, 98.55% (68/69) belonged to Hla sub-geno-type which was the predominant sub-genotype, and only one strain (1.45%) belonged to H1b sub-genotype by RFLP analysis which was in accordance with the results by DNA sequencing method. Phylogenetic tree analysis indicated the H1a sub-genotype measles virus strains should be further divided into 2 clades, and the variation fluctuated between 0.2% and 3.8%. There were transmission chains caused by different virus strains co-cireulation. Conclusion A genotype, H1a and H1b sub-genotype can be identified by RT-PCR-RFLP assay specically based on the restriction enzyme Bcn I .The RT-PCR-RFLP assay can be a rapid, simple, accurate and efficient method for large-scale surveillance of measles virus strains in China.

Objective To measure the level of diamine oxidase (DAO), and observe the intestinal motor and mucosal barrier injury after acute spinal cord injury (SCI) in rats. Methods A total of 45 Sprague-Dawley rats were randomly divided into SCI group (group A, n=15), sham group (group B, n=15) and control group (group C, n=15). SCI model was established with Allen's strike mode (10 g × 25 mm) by striking T10 spinal segment in rats. One day, three days and seven days after SCI, hind limb motor function was assessed with Basso-Beat-tie-Bresnahan (BBB) Scale in each group, the myoelectric slow wave and ileum smooth muscle contractility were measured in rats, ileum tis-sues were tested with HE staining, and the DAO content of serum was tested with ELISA kit. Results One day, three days and seven days af-ter SCI, the BBB score was significantly lower in group A than in groups B and C (P<0.001). One day, three days after SCI, the frequency and amplitude of both slow wave and contractility were lower in group A than in groups B and C (P<0.05);seven days after SCI, there was no significant difference among three groups (P>0.05). Group A showed ileal mucosal edema, lodging, inflammatory cell infiltration, and submucosal gap increase. The Chiu's score of intestinal mucosal injury was higher in group A than in groups B and C (P<0.05), as well as the serum DAO content one day and three days after SCI (P<0.05), and no significant difference was found in the serum DAO content among three groups seven days after SCI (P>0.05). Conclusion Serum DAO content may respond to the intestinal motor function and mu-cosal injury after acute SCI in rats.

Objective To investigate the role of endothelial progenitor cells ( EPCs ) transplantation in rats with sepsis induced by endotoxin ( lipopolysaccharides, LPS ). Methods Sixty clean grade Sprague-Dawley ( SD ) rats with genetic background were divided into three groups according to random number table method:control group, model group, and EPCs transplantation group, with 20 rats in each group. The sepsis model was reproduced by intravenous delivery of LPS 5 mg/kg. Rats in control group were injected with the same amount of normal saline. EPCs were isolated, and cultured and identified were fluorescently labeled with the green fluorescent protein ( GFP ) adenoviral transfection method. The EPC transplantation group was injected with LPS, then a fluorescently labeled EPCs suspension was injected via the tail vein 1 hour later. The expression of fluorescent markers of EPCs was detected with both small animal in vivo imaging instrument and frozen section. Seven days after transplantation, abdominal aorta blood was collected to determine interleukins ( IL-6 and IL-10 ) in peripheral blood with enzyme linked immunosorbent assay ( ELISA ), and the lung, liver, and kidney tissues were harvested, the wet/dry ratio of the lung ( W/D ) was calculated, and hematoxylin and eosin ( HE ) staining was performed to observe, the change in histopathology. Toll-like receptor 4 ( TLR4 ) mRNA expression in lung, liver, and kidney tissues was determined with real-time reverse transcription-polymerase chain reaction ( RT-PCR ). Results The positive rate of EPCs cells with double marking of CD133 and CD34 was 99.0% at the 5th generation of subculture by using flow cytometry. After the transplantation of EPCs labeled with the green fluorescent protein, the appearance of fluorescence indicated that EPCs were mainly localized in the chest, and a stronger fluorescence was observed near the blood vessels. EPCs transplantation could significantly reduce the inflammatory cell infiltration and cell damage in lung, liver, and kidney tissue in septic rats. Compared with control group, the expression of IL-6 and IL-10 in the peripheral blood, W/D ratio, and TLR4 mRNA in lung, liver, and kidney were increased significantly in the model group. Compared with model group, the expressions of IL-6 and IL-10 in the peripheral blood were significantly reduced after EPCs transplantation [ IL-6 (μg/L ):2.127±0.118 vs. 2.664±0.438, IL-10 ( ng/L ): 24.5±3.9 vs. 31.5±3.8, both P < 0.01 ]. EPCs transplantation reduced the W/D ratio of lung, liver and kidney tissues ( lung: 4.68±0.24 vs. 5.48±0.15, liver: 3.33±0.11 vs. 3.94±0.09, kidney: 4.08±0.20 vs. 4.84±0.21, all P < 0.05 ], and down-regulated the expression of TLR4 mRNA ( ×103, lung: 782±131 vs. 1 136±126, liver: 39.1±14.0 vs. 69.2±8.7, kidney: 52.2±15.2 vs. 83.5±17.1, all P < 0.01 ). Conclusions EPCs can enter the lung, liver and kidney tissues of the rat successfully after transplantation of EPCs via vein. EPCs transplantation can down-regulate pro-inflammatory process, help to recover the balance of pro-and anti-inflammatory processes, alleviate the damage to the lung, liver, and kidney tissue significantly.

Objective To explore the effects of repetitive transcranial magnetic stimulation (rTMS) on cognitive ability in patients suffering from mild cognitive impairment (MCI) after ischemic stroke.Methods Forty five ischemic stroke survivors with MCI but not meeting the criterion for diagnosis as dementia were recruited, and were randomly assigned into an rTMS group (32 patients) and a control group (30 patients) according to a random number table.Both groups received the routine drug therapy of medicine and cognitive function training, and the rTMS group was additionally given rTMS over the left dorsolateral prefrontal cortex at 5 Hz and 80％ motor threshold.The treatments lasted for 4 weeks.The Montreal Cognitive Assessment (MoCA) and auditory event related potential (ERP) were tested for both group before and after the treatment.Results After the treatment, two groups showed significant improvements in the average score of MoCA compared to that before the treatment, and that of the rTMS group was significantly higher than that of the control group.For both groups, the P300 latency shortened and the amplitude increased after the treatment.Moreover, the latency and amplitude of the rTMS group increased to 355.67 ± 16.43 ms and 8.69 ± 1.65 μV, respectively, after the treatment, significantly shortened and lengthened than that of the control group [(372.76 ± 23.35 ms and 7.03 ± 3.04 μV), respectively].Conclusions rTMS can significantly improve the cognitive ability of ischemic stroke survivors in a relatively safe way.

AIM:To construct the recombinant eukaryotic expression plasmid pEGFP-C1-SR-A I for the high expression in 293T cells in order to identify functions of savenger receptor-A I(SR-A).METHODS:The primer was designed according to MSR1 cDNA and pEGFP-C1-SR-A I was constructed by standard molecular cloning technique and enzyme digestion.After sequencing,the plasmid was transfected into 293T cells by lipidosome method.The expression of scavenger receptor-A I was identified by RT-PCR and Western blotting.The foam cells were evaluated by the formation of lipid granules in the cells with oil red staining.Cell adhesion was analyzed by cell adhesion assay.RESULTS:24 h after transfection,SR-A I mRNA was highly expressed and the high level of the protein was detected.The ratio of foam cell formation was doubled,the efficacy of cell adhesion was enhanced two times compared to the control group and the empty vector group.CONCLUSION:The recombinant eukaryotic expression plasmid has been constructed successfully with enhancing the function of uptake ox-LDL and adhesion in 293T cells by overexpression of SR-A I.